Characterisation of a new molecule based on two E2 sequences from bovine viral diarrhoea-mucosal disease virus fused to the human immunoglobulin Fc fragment
| Autor: | Alaín González Pose, Rafael Maura Pérez, Claudia Altamirano Gómez, Raquel Montesino Seguí, Jorge R. Toledo, Ignacio Cabezas Ávila, Oliberto Sánchez Ramos, Florence Hugues Salazar |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
e2 glycoprotein
040301 veterinary sciences viruses Veterinary medicine Virus Epitope immune response Viral vector law.invention expression system 0403 veterinary science 03 medical and health sciences Immune system law SF600-1100 bovine viral diarrhoea virus 030304 developmental biology 0303 health sciences General Veterinary biology Chemistry Immunogenicity 04 agricultural and veterinary sciences Molecular biology Cell culture adenoviral vector Recombinant DNA biology.protein Antibody Research Article |
| Zdroj: | Journal of Veterinary Research, Vol 65, Iss 1, Pp 27-37 (2021) Journal of Veterinary Research |
| ISSN: | 2450-8608 |
| Popis: | Introduction Proper conformational arrangement of the E2 molecules of bovine viral diarrhoea-mucosal disease virus (BVD-MDV) is crucial to obtain an effective recombinant vaccine candidate against the disease. In this study, we characterised a new molecule composed of two distinct sequences of the E2 glycoprotein of BVD-MDV and the Fc fragment of human immunoglobulin (BVDE2Fc). Materials and Methods The chimaeric protein was expressed in mammalian cell lines of different species by adenoviral transduction and purified by immobilised metal-affinity chromatography. The N-glycans were profiled by HPLC, and the BVDE2Fc immunogenicity was assessed in male mice. The antigen-antibody reactions were evaluated by ELISA. Results The MDBK cell line was selected from among five for the final production of BVDE2Fc. After purification to over 90%, the N-glycan profile showed neutral and complex oligosaccharides. The mouse immunisation induced a strong humoral response, which produced antibodies able to attach to conformational epitopes on E2 molecules, while the Fc fragment barely contributed to the immune response. Additionally, BVDE2Fc attached to antibodies from bovine sera positive to distinct BVD-MDV subtypes, whereas the loss of BVDE2Fc structure during the deglycosylation process considerably diminished those interactions. Conclusion These results demonstrate that the structure of E2 molecules arranged in tandem and attached to an Fc fragment could represent a viable design for future vaccine candidates against BVD-MD. |
| Databáze: | OpenAIRE |
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